Emma Hanson

Martinson
Advanced Biology
11-19-14
The Effect of Temperature, pH and Concentration on Enzymes
Introduction
Enzymes are classified as highly specific proteins that function as catalysts working
to facilitate biochemical reactions by lowering the activation energy needed for the
reaction. Enzymes consist of two parts, the active site; which binds to the substrate
forming an enzyme-substrate complex that aids in the chemical reaction, and the
allosteric site; which links to specific cofactors forming the correct 3 dimensional
shape. The ruction can be summarized with the equation :( Enzyme + Substrate
Enzyme Substrate Complex Enzyme + Product), you can see by this equation that
the enzymes are not consumed by the reaction; rather they are recycled and
reused. Because these sites are very unique each individual enzyme requires highly
specific terms and conditions of the reactions in which they will catalyze. Because of
how specialized they are enzymes are also easily affected by external factors such
as temperature pH and concentrations.
Methods
a. Materials (for all procedures)

50 mL beaker
containing fresh
catalase (yeast)
solution
Reaction chamber
Ring stand
Clamp

10mL graduated
cylinder
100mL graduated
cylinder
3% hydrogen
Peroxide
Pan (water bath)
Pipette

Hot plate
Ice
Thermometer
Boiled catalase
Buffers with pH: 4, 7
and 10
Distilled water
Balance
NaCL (salt)

b. Procedure Summary of general lab procedure

Begin by using the water pan, ring stand, clamp, 100mL graduated cylinder and
reaction chamber to create a setup as illustrated in the diagram below. Create a
column of water in the graduated cylinder by submerging the cylinder in the water
completely then flip it upside down and secure it to the ring stand being sure to
keep the opening submerged. To test Part A; combine 1mL catalase solution with
10mL hydrogen in the reaction chamber and immediately seal the chamber with the
stopper and hold under water. Measure the level of gas in the cylinder at 30 second
intervals for 5 minutes. For Parts B, D, E and F repeat this test 3-4 times altering the
enzyme concentration, pH, substrate concentration and ionic concentration for each

Hanson 2
respective test. For Part C repeat the test 3 times changing the temperature of the
water for each test.

c. Statistical method Rate

The rate of the reaction quantitatively measured by the difference in each
consecutive recordings of the amount of water displaced in the graduated cylinder
by the oxygen produced in the reaction.

rate( for one interval)=

amount of product 2a mount of product 1

time 2time 1

To find to rate of reaction for each test take the average of the rates for all five
intervals

Average rate=

rate1 +rate2 +rate 3+ rate4 +rate5

5

Part A: Time
a. Data table Team and Class data
Table 1.1: Team Data for the Time Course of Enzyme Activity using 3%
Time

Differen
ce

30se
c
0

1mi
n
0

1:30
0

2mi
n
1

2:30
2

3mi
n
3

3:30
4

4mi
n
6

4:30

5mi
n
6

Table 1.2: Class Data for the Time Course of Enzyme Activity using 3%
The Time Course of Enzyme Activity using 3%
Time

JO,JN

AA,KS,OH

KS,DH,JK,
BE
0

JW,EH,AR

MH,ND,MM

Average

SM,HB,R
W
0

30sec

1.5

3.5

1.333333

1min

2.333333

Hanson 3
1:30:0
0
2 min

3.5

3.5

4.416667

2:30:0
0
3 min

4.5

7.5

5.333333

3:30:0
0
4 min

6.5

10

10

7.333333

4:30:0
0
5min

10.5

11

8.083333

10

12

12

8.833333

b. Graph Team and class data

Graph 1.1: The Time Course of Enzyme Activity using 3%

10
9
8
7
6
Amount of Water Displaced(in mL)

Team Data

Class Data

3
2
1
0

Time Interval

Part B: Enzyme Concentration

a. Data table Team and Class data
Table 2.1: Team Data on the Effect of Enzyme Concentration on Enzyme Activity
Time

30se
c

1mi
n

1:30

2mi
n

2:30

3mi
n

3:30

4mi
n

4:30

5min

Hanson 4

Differen 0
2
2
ce
.50mL Concentration catalase
Differen 0
2
3
ce
.25mL Concentration catalase
Differen 0
1
3
ce

.75mL Concentration catalase

2
3
4
4

Table 2.2: Class Data on the Effect of Enzyme Concentration on Enzyme Activity
Part B:

The Effect of Enzyme Concentration on Enzyme Activity

.75mL Concentration catalase

Time

JOJN

ND MH
MM
0

AA, KS,
OH
0

SM, HB,
RW
0

JW EH
AR
0

Total

KS,DH,J
K,BE
0

30

4.5

15.5

2.583333

2.5

19.5

3.25

1:30:00

3.5

23.5

3.916667

26

4.333333

2:30:00

5.5

29.5

4.916667

6.5

5.5

31

5.166667

3:30:00

34

5.666667

7.5

38.5

6.416667

4:30:00

7.5

7.5

40

6.666667

43

7.166667
Class
Average

Class
average
0

.50 mL Concentration catalase

JOJN

ND MH
MM
0

AA, KS,
OH
0

SM, HB,
RW
0

JW EH
AR
0

Total

KS,DH,J
K,BE
0

30

0.5

9.5

1.5

1.5

1.5

15

2.5

1:30:00

17

2.833333

3.5

20.5

3.416667

2:30:00

2.3

23.3

3.883333

4.5

2.5

26

4.333333

3:30:00

2.5

27.5

4.3

6.5

29.5

4.916667

4:30:00

5.5

30.5

5.083333

5.5

3.5

32

5.333333

Hanson 5
.25mL Concentration catalase
JOJN

ND MH
MM
0

AA, KS,
OH
0

SM, HB,
RW
0

JW EH
AR
0

Total

KS,DH,J
K,BE
0

30

0.5

10.5

1:30:00

0.5

2.5

11

1.1666
67
1.75
1.8333
33
2
2.25
2.5
2.5833
33
2.9166
67
2.9166
67
3.0833
33

12

2:30:00

3.5

13.5

1.5

4.5

15

3:30:00

4.5

15.5

1.5

17.5

4:30:00

1.5

17.5

5.5

18.5

b. Graph Team and Class data

Class
Average

Hanson 6

Graph 2.1 The Effect of Enzyme Concentration on Enzyme Activity .75mL Catalyse Concentration
10
9
8
7
6
Amount of Water Displaced(in mL)

Team Data

Class Data

4
3
2
1
0
Time Interval

Graph 2.2 The Effect of Enzyme Concentration on Enzyme Activity .50mL Catalyse Concentration
8
7
6
5
4
3
2
Team Data
1
Amount of Water Displaced(in mL)
0

Class Data

Time Interval

Hanson 7

Graph 2.3 The Effect of Enzyme Concentration on Enzyme Activity .25mL Catalyse Concentration
4.5
4
3.5
3
2.5

Team Data

Amount of Water Displaced(in mL) 2

Class Data

1.5
1
0.5
0
Time Interval

c. Questions: # 7 parts a-c

Part C: Temperature
a. Data tableTeam and Class data
Table 3: Class Data on the Effect of Temperature on Enzyme Activity
Part C:The Effect of Temperature on Enzyme Activity
5C
Time

SM,
HB,
RW

37C

JOJN

Total

Class
Aver
age
0

0.75

14

12.5

20.5

17.5

25.5

22.5

30.5

24.5

10

34.5

30

1.5

1.5

1:30

2.3

3.3

2.5

4.5

2:30

SM,
HB,
RW

JOJ
N

100C

Total

1
1.65
2.25
3

Clas
s
Aver
age

0
7
10.2
5
12.7
5
15.2
5
17.2
5

SM,
HB,
RW

JOJ
N

Total

Clas
s
Aver
age

0
1.25

0.5

2.5

0.5

2.5

0.5

2.5

0.5

2.5

0.5

2.5

1.25
1.25
1.25
1.25

Hanson 8
3

3.5

6.5

3:30

3.7

6.7

4.5

7.5

4:30

4.9

8.9

5.3

9.3

26.5

10

36.5

29.5

11

40.5

32.5

11

43.5

34.5

12

46.5

35.5

12

47.5

3.25
3.35
3.75
4.45
4.65
b. Graph Class data

18.2
5
20.2
5
21.7
5
23.2
5
23.7
5

1.5

3.5

1.5

3.5

1.5

3.5

1.5

3.5

1.5

3.5

1.75
1.75
1.75
1.75
1.75

Hanson 9

Graph 3 Class Data on The Effect of Temperature on Enzyme Activity

25

20

15

37C
Amount of Water5CDisplaced(in mL) 10

100C

Time Intervals

Hanson 10
c. Questions -- # 8 parts a-c
Part D: pH
a. Data table Team and Class data
Table 4.1 Team Data on the Effect of pH on Enzyme Activity
Time

30se
c

1mi
n

1:30

2mi
n

2:30

3mi
n

.75mL Concentration catalase

Differen 0
3
3
3
4
4
5
ce
.50mL Concentration catalase
Differen 0
1
2
2
3
4
5
ce
.25mL Concentration catalase
Differen 0
2
3
3
4
4
4
ce
Table 4.2 Class Data on the Effect of pH on Enzyme Activity

3:30

4mi
n

4:30

5min

JW
EH
AR
0

Aver
age

Part D:The Effect of pH on Enzyme Activity

pH

Time

ND
MH
MM

AA,K
S,OH

7
JW
AR
EH

Aver
age

ND
MH
MM
0

AA,K
S,OH

10

JW
AR
EH
0

30

3.5

2.66
6667
3.5

1:30

3.5

3.5

3.5

4.5

2:30

3.83
3333
4

5.5

4.5

4.5

3:30

4.5

10

11

4:30

12

4.83
3333
5.33
3333
5.33
3333
5.66
6667

13

b. Graph Team and class data

Aver
age

1.66
6667
2.66
6667
3.33
3333
4.5

2.5

3.5

3.5

3.5

5.16
6667
6.66
6667
7

4.5

3.833
333
4.166
667
4.666
667
4.833
333
5

7.66
6667
8

8.33
3333

5.5

ND
MH
MM
0

AA,
KS,
OH
0

5.333
333
5.333
333
5.833
333

Hanson 11

Graph 4 Team and Class Data on The Effect of pH on Enzyme Activity

9
8
7

Amount of Water Displaced(in mL)

Team Data 4

Team Data 7

Class Average 4
Class Average 4

Team Data 10

Class Average 10

2
1
0
Time Interval

c. Questions --# 9 parts a-c

Part E: Substrate concentration
a. Data table Team and Class data
Part E:The Effect of Substrate Concentration on Enzyme Activity
0%
0.30%
Time

KS,D
H,JK,
BE
0

30

AA,
KS,
OH

MH,
ND,
MM

Aver
age

1.50%

KS,J
K,DH
,BE
0

AA,
KS,
OH
0

MH,
ND,
MM
0

Aver
age

KS,J
K,DH
,BE
0

AA,
KS,
OH
0

MH.
ND.
MM
0

Aver
age

1.666
667
2

2.5

2.5

1:30

4.5

2.333
333
2.5

2:30

5.5

4.25

3.666
667
3.833
333
4

3.5

5.5

3.166

4.5

4.5

Hanson 12
667
3:30

5.5

5.5

4:30

5.5

5.5

3.166
667
4.75

3.833
333
4.75

5.333
333
5.666
667
6
6.333
333

b. Graph Team and class data

Graph 5 : Class Data on The Effect of Substrate Concentration on Enzyme Activity

7
6
5
1.50%

4
Amount of Water Displaced(in mL)

0.30%
3

0%

2
1
0
Time interval

c. Question -- # 10 parts a-c

Part F: Ion concentration
a. Data table Team and Class data
Table 6.1 Team Data on The Effect of Ionic Concentration on Enzyme Activity
Time

30se
c

1mi
n

1:30

2mi
n

2:30

3mi
n

3:30

4mi
n

4:30

5mi
n

Hanson 13

Differen
ce

Differen
ce

Differen
ce

10% NaCl
3
3

3.5

3.5

3.5

3.5

2% NaCl
3
3.5

3.5

4.5

4.5

5.5

5.5

1.5

0% NaCl
2
3

Table 6.2 Class Data on the Effect of Ionic Concentration on Enzyme Activity
Part F:The Effect of Ionic Concentration on Enzyme Activity
10% NaCl
2%
0%
JOJN SM,
Aver
KS,
JOJ
SM,
Aver
KS,
JOJ
SM,
HB,
age
JK,
N
HB,
age
JK,
N
HB,
RW
DH,
RW
DH,
RW
BE
BE
0
0
0
0
0
0
0
0
0
0

Time

KS,J
K,DH
,BE

30

3.666
667

1:30

3.5

4.166
667

2:30

3.5

3.5

3:30

3.5

4.333
333
4.333
333
4.833
333

3.5

4:30

3.5

3.5

Aver
age

2.33
333
3
4

3.5

2.5

1.5

3.5

3.5

3.5

4
4.33
333
3
4.5

3.5

4.5

4.5

4.833
333
4.833
333

4.5

5.5

4.83
333
3
5

3.833
333
4.666
667
5

5.5

4.5

4.833
333

5.5

5.16
666
7
5.16
666
7

5.333
333
5.5

4.5

6.5

Hanson 14
b. Graph Team and class data

Graph 6 Class and Team Data on The Effect of Ionic Concentration on Enzyme Activity
7
6
5
4
3
Amount of Water Displacced( in mL)

10% NaCl Class Data

2% NaCl Class Data

0% NaCl Class Data

10% NaCl Team Data

2% NaCl Team Data

TimeInterval

0% NaCl Team Data